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No Evidence of Genetic Variation in Microsatellite and Mitochondrial Bird Conservation International (2015) 25 :127 –138 . © BirdLife International, 2014 doi:10.1017/S0959270914000203 No evidence of genetic variation in microsatellite and mitochondrial DNA markers among remaining populations of the Strange-tailed Tyrant Alectrurus risora , an endangered grassland species ADRIAN S. DI GIACOMO , ALEJANDRO G. DI GIACOMO , RAFI KLIGER , JUAN C. REBOREDA , RALPH TIEDEMANN and BETTINA MAHLER Summary The Strange-tailed Tyrant Alectrurus risora (Aves: Tyrannidae) is an endemic species of southern South American grasslands that suffered a 90% reduction of its original distribution due to habitat transformation. This has led the species to be classified as globally Vulnerable. By the beginning of the last century, populations were partially migratory and moved south during the breeding season. Currently, the main breeding population inhabits the Iberá wetlands in the province of Corrientes, north-east Argentina, where it is resident all year round. There are two remaining small populations in the province of Formosa, north-east Argentina, and in southern Paraguay, which are separated from the main population by the Parana-Paraguay River and its continuous riverine forest habitat. The populations of Corrientes and Formosa are separated by 300 km and the grasslands between populations are non-continuous due to habitat transformation. We used mtDNA sequences and eight microsatellite loci to test if there were evidences of genetic isolation between Argentinean populations. We found no evidence of genetic structure between populations ( ΦST = 0.004, P = 0.32; F st = 0.01, P = 0.06), which can be explained by either retained ancestral polymorphism or by dispersal between populations. We found no evidence for a recent demographic bottleneck in nuclear loci. Our results indicate that these populations could be managed as a single conservation unit on a regional scale. Conservation actions should be focused on preserving the remaining network of areas with natural grasslands to guarantee reproduction, dispersal and prevent further decline of populations. Resumen El Yetapa de Collar Alectrurus risora (Aves: Tyrannidae) es una especie endémica del cono sur de Sudamérica que ha sufrido una reducción del 90% de su distribución geográfica histórica debido a la transformación del hábitat. Esto condujo a que la especie sea clasificada como vulnerable a nivel global. A principios del siglo pasado, las poblaciones eran parcialmente migratorias migrando al sur durante la temporada reproductiva. Actualmente la principal población reproductiva habita en los Esteros del Iberá en la provincia de Corrientes, en el noreste de Argentina, donde es residente durante todo el año. Existen otras dos pequeñas poblaciones restantes en la provincia de Formosa, en el noreste de Argentina, y en el sur del Paraguay, que están separadas de la población principal por el río Paraná-Paraguay y los hábitats de selva ribereña continua. Las poblaciones de Corrientes y Formosa están separadas por 300 km y los pastizales entre ambas poblaciones no son continuos A. S. Di Giacomo et al. 128 debido a la fragmentación por los cambios recientes en el uso de la tierra. El objetivo de este trabajo fue estudiar la estructura genética entre ambas poblaciones utilizando secuencias de ADN mitocondrial y ocho microsatélites, con el fin de inferir patrones de aislamiento. No se encontraron evidencias de estructura genética entre las poblaciones de Argentina (Φ ST = 0.004, P = 0.32; F st = 0.01, P = 0.06), lo que puede explicarse ya sea por la retención de un polimorfismo ancestral o por la dispersión actual de los individuos entre ambas poblaciones. Tampoco se encontraron evidencias de un cuello de botella demográfico reciente. Estas poblaciones podrían ser manejadas como una única unidad de manejo a escala regional y las acciones de conservación deberían enfocarse en la conservación de una red de áreas con pastizales naturales que garanticen la reproducción, la dispersión y eviten así la disminución de las poblaciones actuales de la especie. Introduction Loss of suitable habitat and the subsequent fragmentation of populations are recognised as the major threat affecting birds (Owens and Bennett 2000 , Fischer and Lindenmayer 2007 , Hoffman et al. 2010 , Rands et al. 2010 ). Among the variety of biological processes negatively affected by habitat fragmentation, one of the most important is the geographic structure of genetic diversity and the reduction in gene flow among populations (Höglund 2009 , Frankham et al. 2010 ). Also, the small population sizes in remaining fragments may cause increased levels of inbreeding and a stronger effect of random genetic drift (Wade 2001 ). Both processes result in a loss of genetic vari- ability within populations, which generally causes a reduction in individual fitness and population adaptability and ultimately may lead to local and global extinction (Gilpin and Soulé 1986 , Hedrick and Kalinowski 2000 , Höglund 2009 ). Loss of gene flow because of reduced dispersal, and reduction in genetic diversity because of genetic drift, can re-distribute genetic variability among spatially structured populations over the course of only few generations (Harrison and Hastings 1996 , Caizergues et al. 2003 ). Patterns of genetic variation can provide insights into dispersal patterns and population connectivity on scales that direct field measures of dispersal are unable to address (Koenig et al. 1996 ). Thus, genetic information, including data on population genetic structure, has become relevant in species conservation (Frankham 2010 , Haig et al. 2011 ). Ryder ( 1986 ) introduced the concept of evolutionarily significant units (ESU) to account for genetic diversity below recognised taxonomic levels. Although ESU definitions might differ (Moritz 1994 , Crandall et al. 2000 , Kizirian and Donnelly 2004 , Degner et al. 2007 ), genetic data always play a critical role for identifying evolutionary units for conservation purposes (for a review see Palsbøll et al. 2006 ). The protection of genetically distinct populations helps to maintain high levels of biodiversity minimising extinction risks (Waples 1991 ). The Strange-tailed Tyrant Alectrurus risora belongs to the group of Tyrant flycatchers (Aves: Tyrannidae), which are endemic of the New World. Thirteen percent of the 429 species of this group (Fitzpatrick 2004 ) are threatened (10 ‘Endangered’ and 19 ‘Vulnerable’ species) or ‘Near Threatened’ (27 species) at global level (Birdlife International 2013a ). Most of the tyrant flycatchers are typical forest inhabitants (Fitzpatrick 1980 , 2004 ), although some of them have diversified towards open habitats occupying grasslands, steppes and wetlands along the Andes, Patagonia and Pampas in western and southern South America (Ohlson et al. 2008 ). Populations of endemic Neotropical grassland birds, including tyrant flycatchers, have declined dramatically over recent decades in these habitats where they now exist mostly in isolated fragments scattered in an agricultural matrix (BirdLife International 2008, Azpiroz et al. 2012 ). The Strange-tailed Tyrant has suffered a large contraction reaching a 90% loss of its original range. This was associated with the conversion of natural grasslands to agriculture during the last century (Di Giacomo and Di Giacomo 2004). At present, it is classified as a globally ‘Vulnerable’ species (IUCN 2013 ) and further decline in range and number is expected because suitable Conservation genetics of the Strange-tailed Tyrant 129 habitats are subject to extensive agricultural modification, intensification of livestock farming and afforestation (Collar et al. 1992 , BirdLife International 2013b). Range contraction was accompa- nied by a change in migration behavior. While one century ago the species was partially migra- tory and reproduction also occurred in Central Argentina, at present all populations are resident and breed in grasslands of Northern Argentina and Paraguay (Di Giacomo and Di Giacomo 2004, Di Giacomo et al. 2011a). The current main breeding population inhabits Iberá wetlands in the province of Corrientes, north-east Argentina, while other small populations occur in the province of Formosa, north-east Argentina, and in southern Paraguay (Birdlife International 2013 b ; Figure 1 ). The Parana-Paraguay River and its continuous riverine forest habitat historically separate these small populations from the main one. With regard the Argentinean populations, they are sepa- rated by 300 km and the grasslands between them are non-continuous due to fragmentation by changes in land use. However, a comprehensive study that has been carried out on the Formosa population for the last 10 years showed that banded fledglings are rarely seen in the population in the following years, suggesting that young individuals might disperse from their natal area (Di Giacomo et al. 2011b ). The aim of this study was to analyse the genetic variability and the genetic structure of the main population and one isolated Argentinean population of the Strange-tailed Tyrant, using both mtDNA and microsatellite molecular markers. These markers differ in mutation rates and mode of inheritance and provide essential information about population history and demographics (Wayne and Morin 2004 ). Current conservation and management efforts are focused on preserving Figure 1. Map showing the present distribution of the Strange-tailed Tyrant (grey area) from Di Giacomo et al . (2011a). Numbers indicate the location of the two populations
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